We used PCR simultaneously detect both HIV 1 Hepatitis DNA by co- amplification using nested HBV primers. The amplified products were detected using labeled primers to internally synthesized regions. Using the HIV-1 and HBV plasmid DNAs as controls, we were able to detect less than 10 copies of each viral genome. Analysis of clinical serum samples from dually infected individuals resulted in positive PCR results with both HBV and HIV probes whereas two antibody/antigen non-reactive specimens were also PCR negative. Thus, we demonstrated the ability to detect multiple viral species in clinical samples in the same PCR reaction. This may prove useful for analysis of samples with limited volumes and to simplify detection.